摘要
目的:观察抗肝纤维化细胞因子白介素10(IL-10)、肝细胞生长因子(HGF)和干扰素γ(IFN-γ)对含不同-509C>T基因型的转化生长因子β_1(TGF-β_1)基因上游启动子转录活性的影响,探讨细胞因子可能的抗肝纤维化机制。方法:以TGF-β_1基因上游5′侧翼区启动子(-1328～+812)含有-509C>T位点的片段作为靶序列,将其与氯霉素乙酰基转移酶(CAT)报告基因组成重组体phTGF2.14C、phTGF2.14T,脂质体法转染至人肝癌细胞系HepG2,应用IL-10(4ng/ml)、HGF (10ng/ml)和IFN-γ(20ng/ml)作用于转染后HepG2细胞,ELISA法测定报告基因的表达。结果:转染phTGF2.14C的HepG2细胞报告基因活性明显高于转染phTGF2.14T者(P<0.01)。IFN-γ对转染phTGF2.14C、phTGF2.14T的HepG2细胞TGF-β_1启动子转录活性均具有明显的抑制作用(P<0.05),HGF对转染phTGF2.14C HepG2细胞的TGF-β_1启动子转录活性具有促进作用(P<0.05),IL-10对两种情况下HepG2细胞TGF-β_1启动子转录活性调控作用均不显著。结论:C等位基因可明显增强TGF-β_1基因启动子转录活性;IFN-γ可能通过抑制TGF-β_1基因启动子转录活性抑制肝纤维化,而HGF、IL-10的抗纤维化作用可能与此途径无关。
Objective：To study the regulatory effects of antifibrotic cytokines, interleukin 10 （IL-10）, hepatocyte growth factor （HGF）, and interferon-gamma （IFN-7） on activity of transforming growth factor-β1 （TGF-β1） gene promoter, so as to assess the antifibrotic mechanism of cytokines. Methods： Sequence - 1328-＋812 of TGF-β1 gene, which contains the -509 C〉 T polymorphism, was selected as putative promoter. The recombinant constructions containing -1328-＋ 812 of TGF-β1 gene and CAT reporter gene （phTGF2.14T, phTGF2.14C） were constructed and transfected into HepG2 cells with liposomal trans- fection method, then the transfected HepG2 cells were treated with IL-10 （ 4 ng/ml）, HGF （ 10 ng/ml） or IFN-7（20 ng/ml）. Reporter gene activity was analyzed by ELISA. Results： Reporter gene activity in cells transfected with phTGF2. 14C was sig- nificantly higher than those transfected with phTGF2.14T 〈P〈0.01）. IFN-）＇ significantly inhibited the reporter gene activity in HepG2 cells transfected with phTGF2.14C or phTGF2.14T（P〈0.05）; HGF significantly increased the reporter gene activity in cells transfected with phTGF2.14C （P〈0.05）. IL-10 had no effects on the activities of cells transfected with phTGF2.14C or phTGF2.14T. Conclusion： C allele at -509 can increase the promoter activity of TGF-β1 gene in HepG2 cells. The antifibrotic effect of IFN-7 might be related to its inhibitory effect on the putative promoter activity of TGF-β1 gene; the antifibrotic effects of HGF and IL-10 may not be through regulation of TGF-beta; gene transcription.
出处
《第二军医大学学报》
CAS
CSCD
北大核心
2007年第12期1292-1295,共4页
Academic Journal of Second Military Medical University
基金
国家自然科学基金(30270605)~~
关键词
多态性
单核苷酸
白细胞介素10
肝细胞生长因子
干扰素Γ
转化生长因子β
转录
遗传
肝硬化
polymorphism, single nucleotide
interleukin-10
hepatocyte growth factor
interferon-gamma
transforming growth factor beta
transcription, genetic
liver cirrhosis
作者简介
侯丽娜，硕士．
Corresponding author. E-mail： gaocf1115@ yahoo.com
